Redox Proteomic Identification of HNE-Bound Mitochondrial Proteins in Cardiac Tissues Reveals a Systemic Effect on Energy Metabolism After Doxorubicin Treatment

Authors

Y. Zhao, University of Kentucky
Sumitra Miriyala, University of KentuckyFollow
L. Miao, University of Kentucky
Mihail I. Mitov, University of KentuckyFollow
David M. Schnell, University of KentuckyFollow
Sanjit Kumar Dhar, University of KentuckyFollow
J. Cai, University of Louisville
J. B. Klein, University of Louisville
Rukhsana Sultana, University of KentuckyFollow
D. Allan Butterfield, University of KentuckyFollow
Mary Vore, University of KentuckyFollow
I. Batinic-Haberle, Duke University
Subbarao Bondada, University of KentuckyFollow
Daret K. St. Clair, University of KentuckyFollow

Abstract

Doxorubicin (DOX), one of the most effective anticancer drugs, is known to generate progressive cardiac damage, which is due, in part, to DOX-induced reactive oxygen species (ROS). The elevated ROS often induce oxidative protein modifications that result in alteration of protein functions. This study demonstrates that the level of proteins adducted by 4-hydroxy-2-nonenal (HNE), a lipid peroxidation product, is significantly increased in mouse heart mitochondria after DOX treatment. A redox proteomics method involving two-dimensional electrophoresis followed by mass spectrometry and investigation of protein databases identified several HNE-modified mitochondrial proteins, which were verified by HNE-specific immunoprecipitation in cardiac mitochondria from the DOX-treated mice. The majority of the identified proteins are related to mitochondrial energy metabolism. These include proteins in the citric acid cycle and electron transport chain. The enzymatic activities of the HNE-adducted proteins were significantly reduced in DOX-treated mice. Consistent with the decline in the function of the HNE-adducted proteins, the respiratory function of cardiac mitochondria as determined by oxygen consumption rate was also significantly reduced after DOX treatment. Treatment with Mn(III) meso-tetrakis(N-n-butoxyethylpyridinium-2-yl)porphyrin, an SOD mimic, averted the doxorubicin-induced mitochondrial dysfunctions as well as the HNE–protein adductions. Together, the results demonstrate that free radical-mediated alteration of energy metabolism is an important mechanism mediating DOX-induced cardiac injury, suggesting that metabolic intervention may represent a novel approach to preventing cardiac injury after chemotherapy.

Document Type

Article

Publication Date

7-2014

Notes/Citation Information

Published in Free Radical Biology and Medicine, v. 72, p. 55-65.

© 2014 Elsevier Inc. All rights reserved.

This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/.

The document available for download is the author's post-peer-review final draft of the article.

Digital Object Identifier (DOI)

https://doi.org/10.1016/j.freeradbiomed.2014.03.001

Funding Information

This work is supported by NIH Grants CA 049797, CA 139843, Cancer Center Support Grant (CCSG) P30CA177558 from NCI and the Edward P. Evans Foundation.

Related Content

Refer to Web version on PubMed Central for supplementary material.

Repository Citation

Zhao, Y.; Miriyala, Sumitra; Miao, L.; Mitov, Mihail I.; Schnell, David M.; Dhar, Sanjit Kumar; Cai, J.; Klein, J. B.; Sultana, Rukhsana; Butterfield, D. Allan; Vore, Mary; Batinic-Haberle, I.; Bondada, Subbarao; and St. Clair, Daret K., "Redox Proteomic Identification of HNE-Bound Mitochondrial Proteins in Cardiac Tissues Reveals a Systemic Effect on Energy Metabolism After Doxorubicin Treatment" (2014). Toxicology and Cancer Biology Faculty Publications. 74.
https://uknowledge.uky.edu/toxicology_facpub/74